ANALYSES OF BIOACCUMULATION OF ASTAXANTHIN IN FENUGREEK SPROUTS

Abstract

In the present study an attempt was made to use the prawn exoskeleton as biofertilizer to
grown fenugreek sprouts. The presences of astaxanthin from the prawn shell and in the
astaxanthin treated group of plants were analysed by TLC. The antioxidant analyses of the
control fenugreek sprouts and the astaxanthin treated groups showed that in the astaxanthin
treated group the antioxidant level was found to increase from 56% to 76%. In spite of the
good growth the total chlorophyll was found to decrease in the astaxanthin treated plant

Full text

ANALYSES OF BIOACCUMULATION OF ASTAXANTHIN IN
Bharathi Ravikrishnan1*.,
Abirami Selvaraj
1
Department of Biotechnology, Guru Nanak College, Chennai
2
Department of Advanced Zoology and Biotechnology
3
G.S.Gill research Institute, Guru Nanak College,
A R T I C L E I N F O
INTRODUCTION
Malnutrition status in India
Malnutrition is a multidimensional phenomenon and one of the
most depressing issues in India. In broad terms, it may be
divided into protein energy malnutrition and micronutrient
deficiency. India has a high prevalence of micronutrient
deficiency-re
lated health risks, which can be improved by food
fortification (Pam et al
., 2014). According to 2015, February
UNICEF report, India has the largest number of underweight
children and the highest number of undernourished population.
The only right solution
for malnutrition and also eradicating
hunger is to increase the crop productivity fortified with
nutrients (Allen, 2006). Micronutrient deficiency results from
inadequate levels of iron, folate, iodine, and various vitamins;
including A, B6, D, and E, in
the body, leading various
metabolic disorders. After the green revolution, definitely in
India the food production has increased tremendously, but,
there is a huge lacuna to fulfill the nutritional status (Arvind,
2013). Thus, there is an increasing demand
for fortifying food
to meet the nutrition requirements is vital.
Biological significance of Fenugreek plants
In many parts of world plant are used as the best source of
medicine for many centuries (Nasroallah
et al
International Journal of Current Advanced Research
ISSN: O: 2319-6475, ISSN: P: 2319-
6505,
Available Online at
www.journalijcar.org
Volume 6; Issue 11; November
2017; Page No.
DOI:
http://dx.doi.org/10.24327/ijcar.2017.
Article History:
Received 17th August, 2017
Received in revised form 25th
September, 2017
Accepted 13th October, 2017
Published online 28th November, 2017
Key words:
Fenugreek, Astaxanthin,
Antioxidant properties.
Copyright©2017 Bharathi Ravikrishnan et al.
This is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
*Corresponding author:
Bharathi Ravikrishnan
Department of Biotechnology, Guru Nanak College,
Chennai-42
ANALYSES OF BIOACCUMULATION OF ASTAXANTHIN IN
FENUGREEK SPROUTS
Abirami Selvaraj
2., Ragunathan
Manickavallii Gurunadhan
Jayanthi Jayaprakash3
Department of Biotechnology, Guru Nanak College, Chennai
-
42
Department of Advanced Zoology and Biotechnology, Guru Nanak College, Chennai
G.S.Gill research Institute, Guru Nanak College,
Chennai-42
A B S T R A C T
In the present study an
attempt was made to use the prawn exoskeleton as biofertilizer to
grown fenugreek sprouts. The presences of astaxanthin from the prawn shell and in the
astaxanthin treated group of plants were analysed by TLC. The antioxidant analyses of the
control fenugr
eek sprouts and the astaxanthin treated groups showed that in the astaxanthin
treated group the antioxidant level was found to increase from 56% to 76%. In spite of the
good growth the total chlorophyll was found to decrease in the astaxanthin treated plan
Malnutrition is a multidimensional phenomenon and one of the
most depressing issues in India. In broad terms, it may be
divided into protein energy malnutrition and micronutrient
deficiency. India has a high prevalence of micronutrient
lated health risks, which can be improved by food
., 2014). According to 2015, February
UNICEF report, India has the largest number of underweight
children and the highest number of undernourished population.
for malnutrition and also eradicating
hunger is to increase the crop productivity fortified with
nutrients (Allen, 2006). Micronutrient deficiency results from
inadequate levels of iron, folate, iodine, and various vitamins;
the body, leading various
metabolic disorders. After the green revolution, definitely in
India the food production has increased tremendously, but,
there is a huge lacuna to fulfill the nutritional status (Arvind,
for fortifying food
Biological significance of Fenugreek plants
In many parts of world plant are used as the best source of
et al
., 2013).
Among them fenugreek is one of the oldest plant which is
commonly used in traditional medicine in many parts of Asia
(Nathiya et al
., 2012). The whole plant of fenugreek e
at the sapling stage and also its seeds are known for their
curative properties (Mullaicharam
properties are associated with the antioxidant properties of
fenugreek (Petropoulos, 2002). Fenugreek is commonly
consumed
in raw form or in form of cooked form or as dry
form in order to cure obesity, cancer, microbial infection and
also known to control diabetics (Hajimehdipoor
is also known to wade of insects (Qureshi
Prawn shell as biofertilizer
When crustacean shell wastes are left to decay on the sea shore
it putrefies and becomes a useless alkaline compound (Prabu
and Natarajan, 2012). But instead, crustacean exoskeleton
waste can be used to develop a wide range of value added
pr
oducts. According to Thirunavukkarasu and Shanmugam
(2009), the crustacean processing industries throughout world
generated 60,000 tonnes of waste every year. According to
Jeyanthi et al
., 2012; when the seeds of tomato, gram and peas
were found to germina
te faster in the presences of prawn shell
powder and the shoot and root length were found to increase
considerably (Jeyanthi et al
., 2015).
International Journal of Current Advanced Research
6505,
Impact Factor: SJIF: 5.995
www.journalijcar.org
2017; Page No.
7626-7629
http://dx.doi.org/10.24327/ijcar.2017.
7629.1194
This is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Bharathi Ravikrishnan
Department of Biotechnology, Guru Nanak College,
ANALYSES OF BIOACCUMULATION OF ASTAXANTHIN IN
Manickavallii Gurunadhan
2 and
42
, Guru Nanak College, Chennai
-42
attempt was made to use the prawn exoskeleton as biofertilizer to
grown fenugreek sprouts. The presences of astaxanthin from the prawn shell and in the
astaxanthin treated group of plants were analysed by TLC. The antioxidant analyses of the
eek sprouts and the astaxanthin treated groups showed that in the astaxanthin
treated group the antioxidant level was found to increase from 56% to 76%. In spite of the
good growth the total chlorophyll was found to decrease in the astaxanthin treated plan
t.
Among them fenugreek is one of the oldest plant which is
commonly used in traditional medicine in many parts of Asia
., 2012). The whole plant of fenugreek e
specially
at the sapling stage and also its seeds are known for their
curative properties (Mullaicharam
et al., 2013). The medicinal
properties are associated with the antioxidant properties of
fenugreek (Petropoulos, 2002). Fenugreek is commonly
in raw form or in form of cooked form or as dry
form in order to cure obesity, cancer, microbial infection and
also known to control diabetics (Hajimehdipoor
et al., 2010). It
is also known to wade of insects (Qureshi
et al., 2005).
When crustacean shell wastes are left to decay on the sea shore
it putrefies and becomes a useless alkaline compound (Prabu
and Natarajan, 2012). But instead, crustacean exoskeleton
waste can be used to develop a wide range of value added
oducts. According to Thirunavukkarasu and Shanmugam
(2009), the crustacean processing industries throughout world
generated 60,000 tonnes of waste every year. According to
., 2012; when the seeds of tomato, gram and peas
te faster in the presences of prawn shell
powder and the shoot and root length were found to increase
., 2015).
Research Article
This is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

International Journal of Current Advanced Research Vol 6, Issue 11, pp 7626-7629, November 2017
7627
METHODOLOGY
Extraction and Characterization of Astaxanthin
The astaxanthin extracted from Fenneropenaeus merguiensis
exoskeleton shell waste was extracted and characterized by
TLC and UV-spectrum as reported by Abirami et al., 2015.
The astaxanthin which was extracted by using acetone as
solvent was subjected to evaporation and the extract was
suspended in DMSO for the plant treatment.
The pigment was characterised by UV Spectroscopy. The
spectrum scan was performed from 470 to 485nm and the peak
was obtained at 480 nm, which confirms the presence of
carotenoid. The concentration of the pigment was calculated
by using the formula given below (Uma Nath et al.,2012).
AST (µg.g-1) = AxDx106
100xGx dxE1%-1cm
Where: AST is concentration of astaxanthin, A is Absorbance,
D is volume of the hexane extract (2ml), G is the weight of the
sample in gms.→3 gms., d is width= 1 cm., E = 2100
Plant Treatment
The treatment of fenugreek sprouts with natural
bioaccumulation of astaxanthin in the sprouts.
Effect of astaxanthin over seed germination
The seeds of fenugreek (methi) were soaked overnight. The
soaked seeds were then tied in small bag of cotton cloth and
were allowed to germinate and one bundle of seeds were
maintained as control sample. During the germination process,
the seeds expect the control ones were treated with 100μl of
DSMO for four days.
Effects of Astaxanthin over sprouting
The fenugreek seeds were sowed and were allowed to
germinate under standard condition. The paper cups were
divided into two batches containing eight paper cups
(Ref.Figure.1). After sprouting the 100μl astaxanthin was
added to the soil of the sprouted plants in the first batch and in
the second batch, 500μl of astaxanthin was treated for four
days continuously. One paper cup in each batch was
maintained as the control, to which astaxanthin was not
treated.
Detection of astaxanthin by Thin Layered
Chromatography(TLC)
After seven days of sprouting and treatment the plants of each
batch with astaxanthin were removed from the soil and were
subjected to washing with running tap water. The whole plants
from each batch were subjected to homogenization with
acetone and a pinch of magnesium sulphate and the content
was incubated at room temperature for 30 minutes in dark. The
upper phase was discarded and the lower phase was used as
the sample for TLC. Silica gel G was used as the stationary
phase and the mobile phase used was Acetone, butanol and
isopropanol in the ratio 5:2:1 with a few drops of distilled
water.
Antioxidant Property Analysis of Fenugreek Sprouts
The antioxidant property of fenugreek plant was estimated by
using DDPH as substrate (Wollgast et al.,2000). The standard
vitamin C was used as control for the analysis of antioxidant
activity of DPPH substrate.
RESULTS
The 13.4μg.gm-1 of astaxanthin was extracted by using as the
acetone as solvent from the exoskeleton of Fenneropenaeus
merguiensis (Fig.1 and 2).
The seeds of fenugreek when treated with astaxanthin, failed to
sprout and the when the sprouts of fenugreek were treated with
astaxanthin, the leaves of fenugreek were found to accumulate
astaxanthin. The batch of leaf which were treated with 100μl
of astaxanthin showed good plant growth and 500μl of
astaxanthin treated plants, exhibited good accumulation of
astaxanthin, but the plant growth was found to be retarded
(Fig.3).
Fig 1 Exoskeleton of Fenneropenaeus merguiensis
Fig 2 Acetone extract of F. merguiensis exoskeleton
Fig 3 Astaxanthin Treated Fenugreek Plants
A. 100μl of astaxanthin treated fenugreek plant
B. 500μl of astaxanthin treated fenugreek plant
Fig 4 TLC analysis of plant pigments
A: Control fenugreek Pigments
B: Acetone extract astaxanthin from Prawn exoskeleton
C: Astaxanthin treated plants (100μl)

Analyses of Bioaccumulation of Astaxanthin in Fenugreek Sprouts
7628
The fenugreek plant treated with astaxanthin and the control
plant on thin layer chromatography of the plant compounds
were analyzed (Fig.4). The antioxidant property of the control
plant and the experimental plant were analysed with DPPH as
the substrate (Table.1).
DISCUSSION
The mainly pharmacological important compounds found in
fenugreek leaves are steroids, polyphenolic compounds, amino
acids and the seeds contains galactomannans, tryptophan,
alkaloids, choline, vitexin, sapogenins, vitamins like A, B1, C
and nicotinic acid (Mehrafarin et al., 2010 and Mohsen et al.,
2012). This plant can be easily cultivated in any climatic
conditions and in any soil profile and it is also known to
improve the nitrogen content of the soil ((Sadeghzadeh-Ahari
et al., 2009). According to Magda, 2017; the germinated
fenugreek seeds when analysed by gas chromatography and
ion-trap mass spectroscopy contains 4.93% astaxanthin and the
antioxidant property was estimated as 56.30%.
In this present study, the astaxanthin treatment at a
concentration of 100 μl, supported for bioaccumulation and
good plant growth. The antioxidant activity of control
fenugreek and treated fenugreek plants were analyzed and the
antioxidant property of astaxanthin treated fenugreek showed
better antioxidant activity than the control plants. Thus by the
treatment of the extracted astaxanthin from the exoskeleton of
Fenneropenaeus merguiensis plays an important role in the
biofortification of the fenugreek sprouts.
CONCLUSION
The fenugreek plants that were treated with astaxanthin which
was extracted from the exoskeleton of F. merguiensis at low
concentration showed good accumulation of astaxanthin and
thereby enhancing antioxidant activity of the fenugreek
sprouts. This biofortified fenugreek could further enhance the
biological application of fenugreek. Thus such research could
help us to reduce the biological waste and also to enhance the
production of crops plant with improvised biopotentials.
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How to cite this article:
Bharathi Ravikrishnan et al (2017) 'Analyses of Bioaccumulation of Astaxanthin in Fenugreek Sprouts ', International
Journal of Current Advanced Research, 06(11), pp. 7626-7629. DOI: http://dx.doi.org/10.24327/ijcar.2017.7629.1194
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